Ix 81 optical microscope

The morphology of ADSCs isolated from well-vascularized and ischemic tissues were examined using an Olympus IX 81 optical microscope. Images were captured at three different locations per culture plate using a Hamamatsu digital camera connected to Slidebook image acquisition software (Slidebook 4.2.0.10, Olympus, Center Valley, PA, USA).

AFM studies were performed on a JPK NanoWizard 4 BioScience AFM (JPK Instruments, Germany) integrated with an iX81 optical microscope (Olympus, Belgium). SH-SY5Y cells were seeded on a glass bottomed dish and cultured at 37 °C in a humidified atmosphere of 5% CO₂ and 95% air. The cells were exposed to 200 μl of the nanoparticle solutions (60 μg ml⁻¹ of GNPs, GNPs–LA, GNPs–α-Syn and the mixture GNPs–LA/GNPs–α-Syn) in the cell growth medium for 24 h and 72 h prior to the AFM measurements. High-resolution topographical images were obtained in Quantitative Imaging mode (QI) using a silicon tip with a nominal radius of 20 nm, spring constant of 0.02 N m⁻¹ and resonance frequency of 7–10 kHz. Cell elasticity measurements were performed in the Force Spectroscopy mode of JPK and spherical tips of 15 μm were used for this purpose. The data were processed by JPK software and the Young's modulus values were extracted using the Hertz model for the spherical indenter. The spring constant of the cantilever was measured before each experiment using the thermal noise method in the JPK software. All AFM studies were performed under physiological conditions at 37 °C and in the appropriate cell medium.

Our setup for collecting the training and testing data for all experiments is composed as a simple reflection mode microscope system (refer to Fig. 1c for the general schematic). We use an Olympus IX81 optical microscope as the centerpiece for our setup. The optical microscope features multiple ports and attachments, including an input reflection mode optical chamber to attach a light source to, and two output chambers that are connected to an attachable camera and optical fiber, respectively. The software and hardware setup allow the optical output of the microscope to be controlled between three paths: the first to the eyepiece for direct visualization, the second to the attached output camera chamber, and the third to the output optical fiber connected to a benchtop spectrometer. In our setup, we used a Hamamatsu ORCA-03G grayscale camera and an Ocean Optics Jaz visual spectrometer at the mentioned output ports. For the incident light, we use a pE-4000 cool LED for selective narrowband illumination, and a halogen lamp (Olympus U-LH100L) coupled to a liquid crystal tunable filter for broadband illumination. Chips with the 1D and 2D grating structures were placed on the microscope stage, with their orientation facing downward for analyzing their reflection mode. A ×4 objective lens (NA = 0.13) was used to analyze the structures and their resonance patterns.